Induction of IFN-β and the Innate Antiviral Response in Myeloid Cells Occurs through an IPS-1-Dependent Signal That Does Not Require IRF-3 and IRF-7

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Abstract
Interferon regulatory factors (IRF)-3 and IRF-7 are master transcriptional factors that regulate type I IFN gene (IFN-α/β) induction and innate immune defenses after virus infection. Prior studies in mice with single deletions of the IRF-3 or IRF-7 genes showed increased vulnerability to West Nile virus (WNV) infection. Whereas mice and cells lacking IRF-7 showed reduced IFN-α levels after WNV infection, those lacking IRF-3 or IRF-7 had relatively normal IFN-b production. Here, we generated IRF-3−/−× IRF-7−/− double knockout (DKO) mice, analyzed WNV pathogenesis, IFN responses, and signaling of innate defenses. Compared to wild type mice, the DKO mice exhibited a blunted but not abrogated systemic IFN response and sustained uncontrolled WNV replication leading to rapid mortality. Ex vivo analysis showed complete ablation of the IFN-α response in DKO fibroblasts, macrophages, dendritic cells, and cortical neurons and a substantial decrease of the IFN-β response in DKO fibroblasts and cortical neurons. In contrast, the IFN-β response was minimally diminished in DKO macrophages and dendritic cells. However, pharmacological inhibition of NF-κB and ATF-2/c-Jun, the two other known components of the IFN-β enhanceosome, strongly reduced IFN-β gene transcription in the DKO dendritic cells. Finally, a genetic deficiency of IPS-1, an adaptor involved in RIG-I- and MDA5-mediated antiviral signaling, completely abolished the IFN-β response after WNV infection. Overall, our experiments suggest that, unlike fibroblasts and cortical neurons, IFN-β gene regulation after WNV infection in myeloid cells is IPS-1-dependent but does not require full occupancy of the IFN-β enhanceosome by canonical constituent transcriptional factors. West Nile virus (WNV) is a mosquito-transmitted virus that infects birds, horses, and humans and has become an emerging infectious disease threat in the Western Hemisphere. In humans, WNV can invade into the brain and spinal cord and destroy neurons, causing severe neurological disease, particularly in the immunocompromised and elderly. A better understanding of how the immune system controls WNV infection is critical for developing new treatments and vaccines. In this study, using a mouse model of WNV infection, we evaluate the combined role of two key transcription factors, interferon-regulatory factor-3 (IRF-3) and IRF-7, that orchestrate antiviral and interferon (IFN) responses after infection. Mice that lack both IRF-3 and IRF-7 were highly vulnerable to lethal infection and cells lacking IRF-3 and IRF-7 had a markedly attenuated IFN-α response. Surprisingly, macrophages and dendritic cells lacking IRF-3 and IRF-7 showed a relatively normal IFN-β response. Furthermore, a genetic deficiency of IPS-1, a protein that signals downstream of the RIG-I- and MDA5 cytoplasmic viral RNA sensors, completely abolished IFN-β production. Our experiments suggest that in specific cell types infected with WNV, IFN-β can be induced through an IPS-1-dependent transcriptional signal that does not require the master transcriptional regulators IRF-3 and IRF-7.